title: Spatial organization of dendritic cells within tumor draining lymph nodes impacts clinical outcome in breast cancer patients.
abstract: BACKGROUND
  Dendritic cells (DCs) are important mediators of anti-tumor immune responses. We hypothesized that an in-depth analysis of dendritic cells and their spatial relationships to each other as well as to other immune cells within tumor draining lymph nodes (TDLNs) could provide a better understanding of immune function and dysregulation in cancer.
  
  
  METHODS
  We analyzed immune cells within TDLNs from 59 breast cancer patients with at least 5 years of clinical follow-up using immunohistochemical staining with a novel quantitative image analysis system. We developed algorithms to analyze spatial distribution patterns of immune cells in cancer versus healthy intra-mammary lymph nodes (HLNs) to derive information about possible mechanisms underlying immune-dysregulation in breast cancer. We used the non-parametric Mann-Whitney test for inter-group comparisons, Wilcoxon Matched-Pairs Signed Ranks test for intra-group comparisons and log-rank (Mantel-Cox) test for Kaplan Maier analyses.
  
  
  RESULTS
  Degree of clustering of DCs (in terms of spatial proximity of the cells to each other) was reduced in TDLNs compared to HLNs. While there were more numerous DC clusters in TDLNs compared to HLNs,DC clusters within TDLNs tended to have fewer member DCs and also consisted of fewer cells displaying the DC maturity marker CD83. The average number of T cells within a standardized radius of a clustered DC was increased compared to that of an unclustered DC, suggesting that DC clustering was associated with T cell interaction. Furthermore, the number of T cells within the radius of a clustered DC was reduced in tumor-positive TDLNs compared to HLNs. Importantly, clinical outcome analysis revealed that DC clustering in tumor-positive TDLNs correlated with the duration of disease-free survival in breast cancer patients.
  
  
  CONCLUSIONS
  These findings are the first to describe the spatial organization of DCs within TDLNs and their association with survival outcome. In addition, we characterized specific changes in number, size, maturity, and T cell co-localization of such clusters. Strategies to enhance DC function in-vivo, including maturation and clustering, may provide additional tools for developing more efficacious DC cancer vaccines.
authors.affiliation: Department of Medicine, Stanford University, 269 Campus Drive, 94305 Stanford, CA, USA. plee@coh.org.
authors.forename: Andrew Y
authors.initials: AY
authors.lastname: Chang
authors.forename: Nupur
authors.initials: N
authors.lastname: Bhattacharya
authors.forename: Jian
authors.initials: J
authors.lastname: Mu
authors.forename: A Francesca
authors.initials: AF
authors.lastname: Setiadi
authors.forename: Valeria
authors.initials: V
authors.lastname: Carcamo-Cavazos
authors.forename: Gerald H
authors.initials: GH
authors.lastname: Lee
authors.forename: Diana L
authors.initials: DL
authors.lastname: Simons
authors.forename: Sina
authors.initials: S
authors.lastname: Yadegarynia
authors.forename: Kaveh
authors.initials: K
authors.lastname: Hemati
authors.forename: Adam
authors.initials: A
authors.lastname: Kapelner
authors.forename: Zheng
authors.initials: Z
authors.lastname: Ming
authors.forename: David N
authors.initials: DN
authors.lastname: Krag
authors.forename: Erich J
authors.initials: EJ
authors.lastname: Schwartz
authors.forename: Danny Z
authors.initials: DZ
authors.lastname: Chen
authors.forename: Peter P
authors.initials: PP
authors.lastname: Lee
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001940:Breast / Q000473:pathology; D001943:Breast Neoplasms / Q000276:immunology* / Q000473:pathology*; D016022:Case-Control Studies; D002449:Cell Aggregation; D002452:Cell Count; D002454:Cell Differentiation; D016000:Cluster Analysis; D003713:Dendritic Cells / Q000276:immunology*; D018572:Disease-Free Survival; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008198:Lymph Nodes / Q000276:immunology* / Q000473:pathology*; D008875:Middle Aged; D013601:T-Lymphocytes / Q000276:immunology; D016896:Treatment Outcome